1. Field of the Invention
The present invention relates in general to a peroxyoxalate chemiluminescence reaction, and more particularly to improvements in luminous time and luminous intensity by adopting an azide compound as a catalyst for the reaction and to various applications utilizing the same.
2. Description of the Prior Art
An analysis method utilizing a chemiluminescene reaction makes it possible to analyze quantitatively all materials which are in relation with a chemiluminescence reaction in a intermediate stage or a last stage of the reaction. It has higher sensitivity than any other than current chemical analysis method and is used in a variety of fields. For example, it is applied for a laboratory analysis test and to a detector of an analyzing equipment. Accordingly, its applicability has attracted attention greatly.
In a chemiluminescence reaction, the reaction associated with luminol, lucigenin, peroxyoxalate and the like has been known in the art. Among these, luminol and peroxyoxalate are representative of the compounds, that are applicable to a practical analysis method.
It has been recognized in the art that a peroxyoxalate chemiluminescence reaction is most effective in the chemiluminescence reactions. As a general reaction path of the peroxyoxalate chemiluminescence reaction, there is an example as follows: ##STR1## wherein F is a fluorescent agent; and F.sup.x is a excited state of F.
In the meantime, considering the peroxyoxalate chemiluminescence reaction, under the existence of a fluorescent agent, an oxalic acid ester compound and a related compound dissolved in a solvent reacts with a peroxide to form 1,4-dioxetane dione, an intermediate, which is then, decomposed by the fluorescent agent to emit light. This chemiluminescence according to the chemiluminescence reaction path shows a relative high efficiency in an alkaline liquid as well as in a neutral liquid and even in a weak acid liquid. As a result, it is widely applied to an analysis test for biological sample such as hydrogen peroxide, peroxygenase, amino acid and the like, as well. In addition, this chemiluminescence reaction method has higher sensitivity than any other quantitative analysis method and has such an advantage that it utilizes various reagents. A variety of analyzing equipments making use of it have been developed. Furthermore, the chemiluminescence reaction is also applied for the principle of a light source and several light sources utilizing it have been already developed and used.
Unlike the chemiluminescence reaction using luminol, however, the chemiluminescence reaction associated with oxalic acid ester compounds and related compounds requires an organic solvent such as an acetonitrile solvent, an ether solvent, an alcohol solvent and the like in order to dissolve the compounds. However, these solvents contain many problems therein such that the compounds have very insignificant solubilities for the solvents and are not stable in the solvents because they decompose spontaneously.
In addition, even oxalic acid ester compound dissolved in a proper solvent, when it is mixed with an aqueous solution in which most analytes are contained, is precipitated because its solubility becomes lowered and thence, it is difficult to induce a complete reaction. Therefore, when the chemiluminescence reaction using oxalic acid ester compounds and related compounds is utilized for a variety of chromatophotometries and other analysis methods, an additional apparatus such as a mixer and the like is needed to prevent such incomplete dissolution and to perform the mixing in harmony. Especially, when the water content of an analyte is high, such solubility decrease prohibits the chemiluminescence analysis method from being applied to the analyte in spite of its high analysis sensisitivity. Furthermore, encountering the analyte with a high water-content, there is always involved a problem in that a luminuous efficiency becomes lowered greatly. The aforementioned problems are serious obstacles to the development of light source for a variety of purposes.
In order to surmount the problems, novel compounds which are capable of being dissolved in water has received careful and much study and investigation. However, the resultant compounds, yet developed has also such problems that most of it is unstable and the luminous flux or intensity thereof is low.
In the meanwhile, several catalysts have been used to increase the luminous intensity and luminous efficiency. They are mostly basic catalysts of which triethylamine and imidazole are representative. However, these basic catalysts can play a role in increasing only reaction rate by changing the hydrogen ion concentration of solution and thus, the aforementioned problems can not be solved thereby. Besides the basic catalysts, metal ions and metal-containing proteins are known as reactive catalysts, but the catalytic effects thereof are unsatisfied and even negligible.